Effect of fixation methods on antibody staining

Core microscopy facilities often receive pre-fixed samples without control over preparation, leading to unpredictable non-specific antibody binding. This study examines how fixation method affects non-specific binding in mouse brain immunohistochemistry. Using 10 brains (5 immersion-fixed, 5 perfusion-fixed), we assessed non-specific antibody staining by counting stained blood vessels. Preliminary results suggest immersion fixation increases non-specific secondary antibody binding. These findings emphasize the need for standardized fixation protocols, as improper fixation can lead to misleading fluorescence signals. Researchers should document fixation methods to ensure accurate staining and imaging results from core facilities.